Allele-specific amplification

• US 9,506,111 B2
• Filed: 10/11/2013
• Issued: 11/29/2016
• Est. Priority Date: 10/20/2008
• Status: Active Grant

• Current Assignee: Roche Molecular Systems (Roche Holding AG)
• Original Assignee: Roche Molecular Systems (Roche Holding AG)
• Inventors: Will, Stephen G., Tsan, Alison, Newton, Nicolas
• Primary Examiner(s): Riley, Jezia
• Application Number:
  US14/052,416
• Publication Number:
  US 20140242592A1
• Time in Patent Office: 1,145 Days
• Field of Search: 536/23.1, 536/24.3, 536/24.33
• US Class Current: 1/1
• CPC Class Codes: C12Q 1/6858 Allele-specific amplification
• CPC Class Codes: C12Q 2525/117 incorporating modified base
• CPC Class Codes: C12Q 2535/125 Allele specific primer extension

Abstract

The present invention includes a method of allele-specific amplification, utilizing an allele-specific oligonucleotide, at least partially complementary to more than one variant of the target sequence, but having a 3′-terminal nucleotide complementary to only one variant of the target sequence and having at least one nucleotide with a base covalently modified at the exocyclic amino group, wherein the allele-specific oligonucleotide is extended by a nucleotide-incorporating biocatalyst predominantly when hybridized to the variant of the target sequence for which it has said complementary 3′-terminal nucleotide.

Claims

1. A kit for allele-specific amplification of a target sequence, which exists in the form of several variant sequences, comprising

(a) a first oligonucleotide, at least partially complementary to one or more variant of the target sequence;

and

(b) a second oligonucleotide, at least partially complementary to one or more variant of the target sequence and having a 3′

-terminal nucleotide complementary to only one variant of the target sequence;

wherein said second oligonucleotide is selected from a group consisting of SEQ ID NOs;

4, 5, 7, 8, 10, 11, and 18.

2. An oligonucleotide for performing an allele-specific amplification of a target sequence, which exists in the form of several variant sequences, comprising

a sequence at least partially complementary to a portion of one or more variants of said target sequence;

a 3′

-terminal nucleotide which is complementary to only one variant of said target sequence and

at least one nucleotide with a base covalently modified at the exocyclic amino group with a sequence selected from a group consisting of SEQ ID NOs;

4, 5, 7, 8, 10, 11 and 18.